LOCUS       V042448                 8218 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042448
VERSION     V042448
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        4..383
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        385..662
                     /label="chicken β-actin promoter"
     intron          663..1680
                     /note="chimera between introns from chicken β-actin and
                     rabbit β-globin"
                     /label="chimeric intron"
     CDS             1741..5199
                     /label="NOS2(NM_000625)"
                     /note="NOS2(NM_000625)"
                     /gene="NOS2"
     CDS             5200..5223
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    5306..5361
                     /note="rabbit&nbsp;β-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
                     /label="β-globin poly(A) signal"
     primer_bind     complement(5722..5738)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5746..5762
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5770..5800)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5814..5835
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        5894..6089
                     /note="SV40 early promoter"
                     /label="SV40 promoter"
     polyA_signal    6095..6229
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(6467..7055)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7226..8086)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8087..8191)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcccc ccccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcggggc gaggcggaaa ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgt tgcgcgctgc cttccccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaaggggc gcggcggccc ccggagcgcc
     1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1381 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1441 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     1561 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1621 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1681 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcgccacc
     1741 atggcctgtc cttggaaatt tctgttcaag accaaattcc accagtatgc aatgaatggg
     1801 gaaaaagaca tcaacaacaa tgtggagaaa gccccctgtg ccacctccag tccagtgaca
     1861 caggatgacc ttcagtatca caacctcagc aagcagcaga atgagtcccc gcagcccctc
     1921 gtggagacgg gaaagaagtc tccagaatct ctggtcaagc tggatgcaac cccattgtcc
     1981 tccccacggc atgtgaggat caaaaactgg ggcagcggga tgactttcca agacacactt
     2041 caccataagg ccaaagggat tttaacttgc aggtccaaat cttgcctggg gtccattatg
     2101 actcccaaaa gtttgaccag aggacccagg gacaagccta cccctccaga tgagcttcta
     2161 cctcaagcta tcgaatttgt caaccaatat tacggctcct tcaaagaggc aaaaatagag
     2221 gaacatctgg ccagggtgga agcggtaaca aaggagatag aaacaacagg aacctaccaa
     2281 ctgacgggag atgagctcat cttcgccacc aagcaggcct ggcgcaatgc cccacgctgc
     2341 attgggagga tccagtggtc caacctgcag gtcttcgatg cccgcagctg ttccactgcc
     2401 cgggaaatgt ttgaacacat ctgcagacac gtgcgttact ccaccaacaa tggcaacatc
     2461 aggtcggcca tcaccgtgtt cccccagcgg agtgatggca agcacgactt ccgggtgtgg
     2521 aatgctcagc tcatccgcta tgctggctac cagatgccag atggcagcat cagaggggac
     2581 cctgccaacg tggaattcac tcagctgtgc atcgacctgg gctggaagcc caagtacggc
     2641 cgcttcgatg tggtccccct ggtcctgcag gccaatggcc gtgaccctga gctcttcgaa
     2701 atcccacctg accttgtgct tgaggtggcc atggaacatc ccaaatacga gtggtttcgg
     2761 gaactggagc taaagtggta cgccctgcct gcagtggcca acatgctgct tgaggtgggc
     2821 ggcctggagt tcccagggtg ccccttcaat ggctggtaca tgggcacaga gatcggagtc
     2881 cgggacttct gtgacgtcca gcgctacaac atcctggagg aagtgggcag gagaatgggc
     2941 ctggaaacgc acaagctggc ctcgctctgg aaagaccagg ctgtcgttga gatcaacatt
     3001 gctgtgctcc atagtttcca gaagcagaat gtgaccatca tggaccacca ctcggctgca
     3061 gaatccttca tgaagtacat gcagaatgaa taccggtccc gtgggggctg cccggcagac
     3121 tggatttggc tggtccctcc catgtctggg agcatcaccc ccgtgtttca ccaggagatg
     3181 ctgaactacg tcctgtcccc tttctactac tatcaggtag aggcctggaa aacccatgtc
     3241 tggcaggacg agaagcggag acccaagaga agagagattc cattgaaagt cttggtcaaa
     3301 gctgtgctct ttgcctgtat gctgatgcgc aagacaatgg cgtcccgagt cagagtcacc
     3361 atcctctttg cgacagagac aggaaaatca gaggcgctgg cctgggacct gggggcctta
     3421 ttcagctgtg ccttcaaccc caaggttgtc tgcatggata agtacaggct gagctgcctg
     3481 gaggaggaac ggctgctgtt ggtggtgacc agtacgtttg gcaatggaga ctgccctggc
     3541 aatggagaga aactgaagaa atcgctcttc atgctgaaag agctcaacaa caaattcagg
     3601 tacgctgtgt ttggcctcgg ctccagcatg taccctcggt tctgcgcctt tgctcatgac
     3661 attgatcaga agctgtccca cctgggggcc tctcagctca ccccgatggg agaaggggat
     3721 gagctcagtg ggcaggagga cgccttccgc agctgggccg tgcaaacctt caaggcagcc
     3781 tgtgagacgt ttgatgtccg aggcaaacag cacattcaga tccccaagct ctacacctcc
     3841 aatgtgacct gggacccgca ccactacagg ctcgtgcagg actcacagcc tttggacctc
     3901 agcaaagccc tcagcagcat gcatgccaag aacgtgttca ccatgaggct caaatctcgg
     3961 cagaatctac aaagtccgac atccagccgt gccaccatcc tggtggaact ctcctgtgag
     4021 gatggccaag gcctgaacta cctgccgggg gagcaccttg gggtttgccc aggcaaccag
     4081 ccggccctgg tccaaggtat cctggagcga gtggtggatg gccccacacc ccaccagaca
     4141 gtgcgcctgg aggccctgga tgagagtggc agctactggg tcagtgacaa gaggctgccc
     4201 ccctgctcac tcagccaggc cctcacctac ttcctggaca tcaccacacc cccaacccag
     4261 ctgctgctcc aaaagctggc ccaggtggcc acagaagagc ctgagagaca gaggctggag
     4321 gccctgtgcc agccctcaga gtacagcaag tggaagttca ccaacagccc cacattcctg
     4381 gaggtgctag aggagttccc gtccctgcgg gtgtctgctg gcttcctgct ttcccagctc
     4441 cccattctga agcccaggtt ctactccatc agctcctccc gggatcacac gcccacagag
     4501 atccacctga ctgtggccgt ggtcacctac cacacccgag atggccaggg tcccctgcac
     4561 cacggcgtct gcagcacatg gctcaacagc ctgaagcccc aagacccagt gccctgcttt
     4621 gtgcggaatg ccagcggctt ccacctcccc gaggatccct cccatccttg catcctcatc
     4681 gggcctggca caggcatcgc gcccttccgc agtttctggc agcaacggct ccatgactcc
     4741 cagcacaagg gagtgcgggg aggccgcatg accttggtgt ttgggtgccg ccgcccagat
     4801 gaggaccaca tctaccagga ggagatgctg gagatggccc agaagggggt gctgcatgcg
     4861 gtgcacacag cctattcccg cctgcctggc aagcccaagg tctatgttca ggacatcctg
     4921 cggcagcagc tggccagcga ggtgctccgt gtgctccaca aggagccagg ccacctctat
     4981 gtttgcgggg atgtgcgcat ggcccgggac gtggcccaca ccctgaagca gctggtggct
     5041 gccaagctga aattgaatga ggagcaggtc gaggactatt tctttcagct caagagccag
     5101 aagcgctatc acgaagatat ctttggtgct gtatttcctt acgaggcgaa gaaggacagg
     5161 gtggcggtgc agcccagcag cctggagatg tcagcgctcg actacaagga cgacgatgac
     5221 aagtagctcg agctagcaga tctttttccc tctgccaaaa attatgggga catcatgaag
     5281 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     5341 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     5401 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     5461 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     5521 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     5581 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     5641 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag
     5701 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5761 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctgggtgc ctaatgagtg
     5821 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5881 tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     5941 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6001 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     6061 gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg
     6121 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6181 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa
     6241 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6301 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6361 gcggtaatac ggttatccac agaatcaggg ataacgcagg aaagaacatg tgagcaaaag
     6421 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6481 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6541 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6601 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6661 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6721 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6781 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6841 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6901 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6961 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7021 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7081 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7141 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7201 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7261 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7321 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7381 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7441 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7501 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7561 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7621 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7681 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7741 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7801 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7861 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7921 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7981 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8041 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     8101 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     8161 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgg
//
